Phloem transport of abscisic acid in Ricinus communis L. seedlings

Sieve tube sap exuded from the cut hypocotyl of castor bean seedlings (Ricinus communis L.) was found to contain 0.2–0.5 mmol m^?3abscisic acid (ABA). The ABA concentration in the sieve tube sap always exceeded that in root pressure exudate under a wide range of water supply. Exudation of sieve tube sap from the cut hypocotyls caused water loss, and this induced ‘water shortage’ in the cotyledons which resulted in the ABA concentration in the cotyledons increasing by 3-fold and that in the sieve tube sap increasing by up to 50-fold within 7h. The wounded surface of the cut hypocotyl was not responsible for the ABA increase. Incubation of the cotyledons of endosperm-free seedlings in various ABA concentrations (up to 100 mmol m^?3) increased the ABA concentration in sieve tube sap. The concomitant increase in ABA, both in cotyledons and in sieve tube sap, had no effect on the phloem loading of sucrose, K⁺ and Mg²⁺ within the experimental period, i.e. up to 10h. It can be concluded that (i) the phloem is an important transport path for ABA, (ii) water stress at the phloem loading sites elevates phloem-mobile ABA, which may then serve as a water stress signal for sinks, for example stem and roots (not only for stomata), and (iii) the ABA concentration of cells next to or in the phloem is more important than the average ABA content in the whole cotyledon for determining the ABA concentration in sieve tube sap.     

Foliar application of nitrate or ammonium as sole nitrogen supply in Ricinus communis. II. The flows of cations, chloride and abscisic acid

Following a precultivation with pedospheric nitrogen nutrition, Ricinus plants were supplied with nitrogen solely by spraying nitrate or ammonium solution onto the leaves during the experimental period. The chemical composition of tissues, xylem and phloem exudates was determined and on the basis of the previously determined nitrogen flows (Peuke et al., New Phytologist (1998), 138 , 657–687) the flows of potassium, sodium, magnesium, calcium, chloride and ABA were modelled. These data, which permit quantification of net-uptake, transport in xylem and phloem, and utilization in shoot and root, were compared with results obtained in plants with pedospherically-supplied nitrate or ammonium and data in the literature. Although the overall effects on the chemical composition of supplying ammonium to the leaves were not as pronounced as in pedospherically supplied plants, there were some typical responses of plants fed with ammonium (ammonium syndrome). In particular, in ammonium-sprayed plants uptake and transport of magnesium decreased and chloride uptake was increased compared with nitrate-sprayed plants. Furthermore, acropetal ABA transport in the xylem in ammonium-sprayed Ricinus was threefold higher than in nitrate-sprayed plants. Additionally, concentrations of anions were more or less increased in tissues, particularly in the roots, and transport fluids. The overall signal from ammonium-sprayed leaves without a direct effect of ammonium ions on uptake and transport systems in the root is discussed.     

Effects of temperature and dinitrophenol on the uptake of potassium and sodium ions in Ricinus communis roots

Summary Detopped root systems of Ricinus communis plants were used for the study of the effects of temperature and DNP on the uptake of K and Na ions supplied as KNO₃ and NaNO₃.When K and Na ions were offered together in equivalent concentrations, the steady state uptake rates for K⁺ and Na⁺ at 23 to 25° gave a K⁺/Na⁺ ratio of 3. Increasing the Na⁺ concentration relative to K⁺ 3-fold did not alter the preferential uptake of K⁺. The uptake of K⁺ was more sensitive to temperature in the range 10 to 40° and to the application of DNP at 1.5x10^-4 M than was the uptake of Na⁺. When NaNO₃ was the only salt supplied Na⁺ uptake became more sensitive to DNP than when both K⁺ and Na⁺ nitrates were supplied. Prolonged application of DNP led to net K⁺ efflux from the roots, even when no K⁺ was being supplied to the roots. Net Na⁺ efflux under the influence of DNP occurred only in roots previously grown on Na-containing nutrient medium.The different responses of the K⁺ and Na⁺ uptake processes to temperature and DNP suggest the operation of different uptake mechanisms for K⁺ and Na⁺ These results have been considered in relation to the recent concept of dual mechanisms for the absorption of alkali cations by plant tissues.     

Effect of moisture stress on abscisic acid levels in Ricinus communis L. with particular reference to phloem exudate

Summary Abscisic acid (ABA) increased considerably in stem apices attached to Ricinus plants during a moisture stress but only slightly in stem apices which were detached and then allowed to lose water. A very marked rise in ABA also occurred in phloem-sap of moisture-stressed Ricinus plants. These results suggest that in apices of moisture-stressed plants the increased ABA does not arise in situ but is translocated to the apex after synthesis in the mature leaves.     

Measurements of pH, sucrose and potassium ions in the phloem sap of castor bean (Ricinus communis) plants

The composition of phloem sap, sampled at different heights along, the stem of castor bean ( Ricinus communis L. cv. Gibsonii) plants, was determined. A gradient in pH was observed; the highest pH values occurred near the shoot apex, decreasing towards the base of the stem. The sucrose content of the exudate exhibited a similar gradient. The concentration of potassium ions was highest near the uppermost, full-grown leaves, decreasing towards the apex and the base of the stem. The importance of these findings for the understanding of phloem translocation and unloading is discussed.     

Delivery rates of abscisic acid in xylem sap of Ricinus communis L. plants subjected to part-drying of the soil

Abscisic acid (ABA) moving from roots to shoots in the transpirationstream is a potential hormonal message integrating perceptionof a root stress with adaptive changes in the shoot. A twinroot system was used to study ways of estimating effects ofdroughting the upper roots of Ricinus communis L. on abscisicacid (ABA) transport to the shoot in the transpiration stream.Droughted plants transpired more slowly than controls. Droughtingalso increased concentrations of ABA up to I I-fold in sap inducedto flow from the roots of freshly decapitated plants at ratesof whole plant transpiration. However, because of dilution effectsarising from the different sap flows in control and droughtedplants, these changes in ABA concentration in the xylem sapdid not accurately reflect amounts of ABA transported. To overcomethis problem, delivery rates were calculated by multiplyingconcentration with sap flow rate to generate ABA delivery interms of µmol s–1 per plant. Droughting for 24 hor more increased ABA delivery from roots to shoots by 5-fold.Since droughting can alter the relative sizes of the roots andshoots and also the root:shoot ratio these delivery rates wererefined in several ways to reflect both the amount of root generatingthe ABA message and the size of the recipient shoot system. Key words: Abscisic acid, Ricinus communis L., soil drying, xylem sap     

The uptake and flow of C, N and ions between roots and shoots in Ricinus communis L. III. Long-distance transport of abscisic acid depending on nitrogen nutrition and salt stress

Seedlings of Ricinus communis L. were cultivated in quartz sandand supplied with media which contained either different concentrationsof nitrate or ammonium nitrogen and were treated with a lowsalt stress. The concentration of ABA was determined in tissuesand in xylem and phloem saps. Between 41 and 51 day after sowing,abscisic acid (ABA) flows between roots and shoots were modelled.Long-distance transport of ABA was not stimulated under conditionsof nitrate deficiency (0.2 mol m^–3). However, when ammoniumwas given as the only N source (1.0 mol m^–3), ABA transportin both xylem and phloem was increased significantly. Mild saltstress (40 mol m^–3 NaCl) increased ABA transport in nitrate-fedplants, but not in ammonium-fed plants. The leaf conductancewas lowered by salt treatment with both nitrogen sources, butit was always lower in ammonium-fed compared to nitrate-fedplants. A negative correlation of leaf conductance to ABA levelsin leaves or flow in xylem was found only in comparison of ammonium-fedto nitrate-fed plants. Key words: Abscisic acid, ammonium, Ricinus communis, phloem, xylem, transport, nitrate, nitrogen nutrition     

Transport of magnesium ions in the phloem of Ricinus communis L. seedlings

During growth of Ricinus communis seedlings, magnesium ions are mobilized in the endosperm, taken up by and accumulated to very high levels (150 μmol·g FW^?1) in the cotyledons, and translocated to hypocotyl and roots. The magnesium gain from days 6 to 7 in the cotyledons and the seedling axis necessitates a total up-take rate of 600 nmol·h^?1-seedling^?1 and the phloem translocation rate must amount to 200 nmol·h^?1. seedling^?1. The phloem loading of magnesium and the regulatory properties of this process were investigated, making specific use of the ability to collect pure phloem sap from the cut hypocotyl of 6-d-old Ricinus seedlings. The concentration of magnesium in sieve-tube sap (5 mM) was fairly constant under many incubation conditions, e.g. incubation in magnesium-free buffer, incubation with different cations (K⁺, Na⁺, NH ₄ ⁺ ) or anions (Cl^?, NO ₄ ^- , SO ₄ ^2- ), or incubation with sucrose and amino acids. Even addition of magnesium chloride to the cotyledons did not enhance phloem loading of magnesium ions. Therefore the high magnesium content of the cotyledons was sufficient for continuous phloem loading of magnesium, irrespective of external ionic conditions. Also, the flow rate of sieve-tube sap did not influence the magnesium concentration in the sap. Only the incubation with sulfate and phosphate ions increased the magnesium-ion concentration in the phloem. Magnesium sulfate offered to the cotyledons caused a threefold increase of magnesium ions in the sieve-tube sap, which was inhibited by Na⁺, NH ₄ ⁺ and Ca²⁺ in rising order, but not by K⁺. Incubation with phosphate for a prolonged period (8 h) led to an increased mobilization of intra-cotyle-donary magnesium and an enhanced phloem loading of mobilized magnesium. It is concluded that phosphate availability is a decisive factor for mobilization and translocation of magnesium ions within the plant.     

The uptake and flow of C, N and ions between roots and shoots in Ricinus communis L. II. Grown with low or high nitrate supply

Seedlings of Ricinus communis L. cultivated in quartz sand weresupplied with a nutrient solution containing either 0.2 molm^–3     

Transport, synthesis and catabolism of abscisic acid (ABA) in intact plants of castor bean (Ricinus communis L.) under phosphate deficiency and moderate salinity

Flows of abscisic acid (ABA) were investigated in whole plantsof castor bean (Ricinus communis) grown in sand culture undereither phosphate deficiency or moderate salinity. Xylem transportof ABA in P-deficient plants was stimulated by a factor of 6whereas phloem transport was affected only very slightly. ABAdeposition into leaves of P-deficient plants was not appreciablydifferent from the controls because of strong net degradationin leaves. Since conjugation of ABA was strongly reduced inall organs of P-deficient plants ABA was presumably metabolizedmainly to phaseic acid and dihydrophaseic acid. The increasedimport of ABA occurred predominantly into fully differentiatedbut not senescent leaves and showed a good correlation withthe inhibition of leaf conductance under P deficiency. As with low-P-plants salt stress increased ABA synthesis inroots and associated transport in the xylem. However, salinitycaused a distinctly greater accumulation of ABA in the leaves,stem segments and the apex than in P-deficient plants. As opposedto P deficiency, ABA export in the phloem from the leaves wasstimulated by salinity. Modelling of ABA flows within an individualleaf over its life cycle showed that young growing leaves importedABA from both phloem and xylem, whereas the adult non-senescentleaves were a source of ABA and thus provided a potential shoot-to-rootstress signal as well as an acceptor for reciprocal signalsfrom root to shoot. In senescing leaves ABA flows and accumulationwere somewhat retarded and ABA was lost in net terms by exportfrom the leaf. Key words: Abscisic acid, phosphorus deficiency, salt stress, phloem and xylem transport     

Endogenous abscisic acid signaling towards storage reserve filling in developing seed tissues of castor bean (Ricinus communis L.)

Abscisic acid (ABA; free form) is a naturally occurring physiological growth hormone of higher plants. A detailed study involving the time course growth of developing seed tissues associated with endogenous levels of free ABA were investigated using a novel enzyme-linked immunosorbent assay. Seed filling in castor (Ricinuc communis L.) endosperm, embryo, and pod is marked with a rapid increase in fresh weight during the mid-developmental stages [21–42 days after pollination (DAP)], followed by a steady decline at the maturation stages (42–63 DAP) accompanied with a rapid lipid synthesis (in endosperm and embryo) during the same period, except for in pod. Endogenous ABA levels in endosperm (0.001–0.32 μg/g) and embryo (0.003–0.13 μg/g) followed a concurrent pattern with seed reserve filling, showing a rapid increase during the mid-developmental stages 21–42 DAP, whereas ABA levels in seed pod (0.2–22.9 μg/g) showed a different accumulation pattern with rapid increase and decline during the early-mid developmental stages, preceded by the maximal increase during the maturation stage (63 DAP). Together, our results provide evidence for the association of endogenous ABA in seed filling as well as in reserve deposition and provides clue for the effective usage of exogenous ABA concentrations in developing seeds with a focus, on improving seed reserve complex in castor.     

Structure and stability of Ricinus communis haemagglutinin.

The molecular properties of the haemagglutinin of Ricinus communis (RCA I or RCA 120) were evaluated by analytical ultracentrifugation, light-scattering, c.d. and fluorescence. The native molecule had a fairly expanded structure (f/f0 = 1.43) and dissociated into two subunits of equal size in 6 M-guanidinium chloride. This native structure was stable in alkali (up to pH 11) and resistant to thermal denaturation at neutrality. A pH-triggered change in the haemagglutinin conformation was observed and characterized by analytical ultracentrifugation, c.d. and fluorescence between pH 7 and 4.5, the range in which its affinity for galactosides decreased [Yamasaki, Absar & Funatsu (1985) Biochim, Biophys. Acta 828, 155-161]. These results are discussed in relation to those reported in the literature for other lectins and more especially ricin, for which a pH-dependent conformation transition has been observed in the same range of low pH.     

Inorganic ions modulate the path of phloem loading of sucrose in Ricinus communis L. seedlings

The impact of inorganic ions on sucrose fluxes in the cotyledons and on the pathway of phloem loading was studied in Ricinus communis L. seedlings. The cotyledons were incubated in defined solutions which contained either potassium, sodium, magnesium or calcium as chloride salts, or the sodium salts of sulphate or phosphate. Sucrose uptake from the medium into the cotyledons was only slightly affected by the salts. Sucrose efflux to the medium was increased by phosphate and sulphate and to a lesser extent by sodium and potassium. Phloem loading from the apoplasm and the symplasm was analysed by addition of labelled sucrose to the medium, determination of the specific radioactivity of sucrose in sieve-tube exudate and quantification of export into the seedling axis. Potassium and sodium stimulated the apoplasmic route of phloem loading of sucrose, mostly at the expense of loading from cotyledon sucrose pools. In contrast, sulphate and phosphate strongly inhibited the apoplasmic route whereas the (small) symplasmic flux from the cotyledon sucrose pools was less affected. Magnesium ions inhibited phloem loading by both pathways. The potential of ions in modulating the pathways of sucrose export in day to day operation of plants is discussed.     

Water dependent and water independent fluxes of potassium in exuding root systems of Ricinus communis

Summary The flux of water, , to the xylem of exuding root systems of Ricinus communis was controlled using a range of mannitol concentrations permitting the influence of this water flux on the potassium flux, f _K, to be studied. The relationship between and f _K thus obtained was investigated, for a number of external concentrations of potassium, Cm, supplied as potassium nitrate. An analysis of these data indicated the presence of a water dependent and a water independent f _K both of which varied with Cm. The water dependent f _K shows a parabolic relationship with Cm for Cm values <1 mM followed by a sharp inflection and decline at higher Cm values whereas the water independent f _K shows an hyperbolic relationship over the same range of Cm values.Uptake of potassium by exuding root systems was measured and shown to be dependent on the solute potential of the medium. The uptake was also shown to exhibit a dual absorption isotherm the kinetics of which indicate a low Km system (system 1) and a high Km system (system 2). The Km value obtained for system 1 is very similar to that obtained for the water independent f _K. It is postulated that the water independent f _K is contributed by that portion of f _K arriving in the stele via the cortical symplast and is directly dependent on Cm. The water dependent f _K is contributed by those ions moved across the root in response to centripetal water movement through the cortical cell walls.     

Lectin affinity high-performance liquid chromatography. Interactions of N-glycanase-released oligosaccharides with Ricinus communis agglutinin I and Ricinus communis agglutinin II

The structural determinants required for interaction of oligosaccharides with Ricinus communis agglutinin I (RCAI) and Ricinus communis agglutinin II (RCAII) have been studied by lectin affinity high-performance liquid chromatography (HPLC). Homogeneous oligosaccharides of known structure, purified following release from Asn with N-glycanase and reduction with NaBH4, were tested for their ability to interact with columns of silica-bound RCAI and RCAII. The characteristic elution position obtained for each oligosaccharide was reproducible and correlated with specific structural features. RCAI binds oligosaccharides bearing terminal beta 1,4-linked Gal but not those containing terminal beta 1,4-linked GalNAc. In contrast, RCAII binds structures with either terminal beta 1,4-linked Gal or beta 1,4-linked GalNAc. Both lectins display a greater affinity for structures with terminal beta 1,4-rather than beta 1,3-linked Gal, although RCAII interacts more strongly than RCAI with oligosaccharides containing terminal beta 1,3-linked Gal. Whereas terminal alpha 2,6-linked sialic acid partially inhibits oligosaccharide-RCAI interaction, terminal alpha 2,3-linked sialic acid abolishes interaction with the lectin. In contrast, alpha 2,3- and alpha 2,6-linked sialic acid equally inhibit but do not abolish oligosaccharide interaction with RCAII. RCAI and RCAII discriminate between N-acetyllactosamine-type branches arising from different core Man residues of dibranched complex-type oligosaccharides; RCAI has a preference for the branch attached to the alpha 1,3-linked core Man and RCAII has a preference for the branch attached to the alpha 1,6-linked core Man. RCAII but not RCAI interacts with certain di- and tribranched oligosaccharides devoid of either Gal or GalNAc but bearing terminal GlcNAc, indicating an important role for GlcNAc in RCAII interaction. These findings suggest that N-acetyllactosamine is the primary feature required for oligosaccharide recognition by both RCAI and RCAII but that lectin interaction is strongly modulated by other structural features. Thus, the oligosaccharide specificities of RCAI and RCAII are distinct, depending on many different structural features including terminal sugar moieties, peripheral branching pattern, and sugar linkages.     

Induced leaf intercellular CO2, photosynthesis, potassium and nitrate retention and strawberry early fruit formation under macronutrient limitation

Relationships between induced high leaf intercellular CO₂ concentrations, leaf K⁺ and NO₃ ^? ion movement and early fruit formation under macronutrient limitation are not well understood. We examined the effects and interactions of reduced K/N input treatments on leaf intercellular CO₂, photosynthesis rate, carboxylation and water use efficiency, berry formation as well as leaf/fruit K⁺, NO₃ ^? and photosynthate retention of strawberry (Fragaria × ananassa Duch.) to enhance low-input agriculture. The field study was conducted in Nova Scotia, eastern Canada during 2009–2010. The experimental treatments consisted of five K₂O rates (0, 6, 12, 18, and 24 kg ha^?1) and five N rates (0, 5, 10, 15, and 20 kg ha^?1), representing respectively, 0, 25, 50, 75, and 100 % of regular macronutrient recommendations based on the soil testing. The treatments were arranged in a split-plot design with three blocks in the field. The cultivar was ‘Mira’, a June-bearing crop. The results showed that strawberry plants treated with 25 %-reduced inputs could induce significantly higher leaf intercellular CO₂ concentrations to improve plant photosynthesis, carboxylation and water use efficiency and translocation of leaf/fruit K⁺ and dissolved solids, which could advance berry formation by 6 days and produce significantly higher marketable yields (P < 0.05). Higher leaf intercellular CO₂ inhibited leaf/fruit NO₃ ^? ion retention, but this inhibition did not occur in leaf/fruit K⁺ retention. Linear interactions of the K/N treatments were significant on fruit marketable yields, intercellular CO₂, net photosynthesis, leaf transpiration rates, and leaf temperatures (P < 0.05). It was concluded that higher leaf CO₂ could enhance plant photosynthesis, promote plant carboxylation and water use efficiency, and advance berry formation, but it could inhibit leaf NO₃ ^? retention. This inhibition did not find in leaf K⁺ ion and dissolved solid retention. Overlay co-limitation of leaf intercellular CO₂ and translocation of leaf/fruit K⁺/NO₃ ^? and total dissolved solids could constrain more fruit formation attributes under full macronutrient supply than reduced inputs. It was suggested that low input would be an optimal and sustainable option for improving small fruit crop physiological development and dealing with macronutrient deficiency challenge.     

Transfer of amino acids and nitrate from the roots into the xylem of Ricinus communis seedlings

The loading of amino acids and nitrate into the xylem was investigated by collection and analysis of root-pressure exudate from the cut hypocotyl stumps of seedlings of Ricinus communis L. Glutamine was found to be the dominant amino acid in the exudate and also to be the amino acid which is transferred to the xylem most rapidly and accumulated to the greatest extent. The comparison between uptake and xylem loading showed significant differences in specificity between these two transport reactions, indicating a different set of transport systems. Nitrate is transferred to the xylem at a higher relative rate than any amino acid despite the great nitrate-storage capacity of the root system. Thus the supply of nitrate to Ricinus plants leads to enhanced nitrogen allocation to the shoots.